Plant Parasitic Nematodes Associated with Jatropha Curcas

PLANT PARASITIC NEMATODES ASSOCIATED WITH JATROPHA CURCAS

ACCESSIONS IN SOME LOCAL GOVERNMENT AREAS OF KADUNA

STATE, NIGERIA

Jeremiah Olamide OLATUNJI, B. AGRIC. (FUNAAB) 2009

(M.SC/AGRIC./15143/2011-2012)

A DISSERTATION SUBMITTED TO THE SCHOOL OF POSTGRADUATE STUDIES, AHMADU BELLO UNIVERSITY, ZARIA IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE AWARD OF A MASTER OF SCIENCE DEGREE IN CROP PROTECTION

DEPARTMENT OF CROP PROTECTION, FACULTY OF AGRICULTURE, AHMADU BELLO UNIVERSITY, ZARIA, NIGERIA.

FEBRUARY, 2016

DECLARATION

I hereby declare that the work in this dissertation titled „„Plant Parasitic Nematodes Associated with Jatropha curcas Accessions in Some Local Government Areas of Kaduna State, Nigeria‟‟ has been performed by me in the Department of Crop Protection under the supervision of Drs. Chindo, P. S., Alao, S. E. L. and. Agbenin, N. O. The information derived from the literature has been duly acknowledged in the text and in the references. No part of this dissertation was previously presented for another degree at any University.

Jeremiah Olamide OLATUNJI ______

Name of Student Signature Date

CERTIFICATION

This dissertation entitled PLANT PARASITIC NEMATODES ASSOCIATED WITH JATROPHA CURCAS ACCESSIONS IN SOME LOCAL GOVERNMENT AREAS OF KADUNA STATE, NIGERIA by Jeremiah Olamide OLATUNJI meets the regulations governing the award of the degree of Master of Science in Ahmadu Bello University and is approved for its contribution to knowledge and literary presentation.

Dr. P. S. Chindo ______(Chairman, Supervisory Committee) Signature Date

Dr. S. E. L. Alao ______(Member, Supervisory Committee) Signature Date

Dr. N. O. Agbenin ______(Member, Supervisory Committee) Signature Date

Prof. A. B. Zarafi ______(Head, Department of Crop Protection) Signature Date

Prof. K. Bala ______(Dean, School of Postgraduate Studies.) Signature Date

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DEDICATION

This work is dedicated to the glory of God Almighty, the author and finisher of my faith and to the memory of my late father, Elder Abel Olabisi Olatunji.

ACKNOWLEDGEMENT

With profound gratitude to the Almighty, all sufficient God with whom all things are possible and whose mercies endureth forever. I express my sincere gratitude to my supervisors, Drs. Chindo, P. S., Alao, S. E. L. and Agbenin, N. O from whose pool of knowledge I have been greatly priviledged to learn. My appreciation goes to the Head of Department, Prof. A. B. Zarafi, for providing conducive environment during my studies. My appreciation goes to lecturers in the Department, technical Staff and Mr. I. Donald. I remain grateful to Prof. C. A. Ichekwu, for provision of Jatropha curcas seeds, the National Research Institute for Chemical Technology (NARICT), Basawa, Zaria for allowing me to visit and take samples from their plantation and eHealth System Africa, Kano, for granting me permission to complete the studies.

“He who has a father will always go farther, and with a mother you will never be murdered” I thank God for giving me parents like you, Late Elder A. O. Olatunji and Deaconess A. O. Olatunji. You will live long and eat the fruit of your labour in Jesus name (Amen). I am also greatly indebted to my loving and wonderful uncle,Mr. S. O. Atsiangbe and my siblings, Engr. and Mrs. S. O. Olatunji, Mr. and Mrs. S. O.Ilo, Mr. and Mrs. Olatunbosun Olatunji and Olayinka Olatunji, I will forever love and cherish you. To my colleagues and friends, Mr. and Mrs. E. O. Ooju, Messers A. Peter, Y. Lurwanu, O. Anifowose and A. Abdullahi, we are the best combination and meet you at the top in the nearest future. I also wish to appreciate Miss I. Q. Amadi for her love and prayers.

ABSTRACT

A study was conducted to determine the genera, frequency and prominence value of plant parasitic nematodes associated with Jatropha curcas accessions in Sabon Gari,

Kudan, Giwa and Zaria Local Government Areas of Kaduna State, Nigeria in 2013.

Using systematic random method of sampling, 72 soil and root samples were collected from the Jatropha plant stands, and nematodes extracted from the samples using the sieving and decanting and modified Baermann pan methods. Twenty-four genera of plant-parasitic nematodes were recorded in all the locations. Plant-parasitic nematodes recovered includes Scutellonema, Hoplolaimus, Pratylenchus, Aphelenchus,

Meloidogyne, Tylenchoryhnchus, Rotylenchus, Longidorus, Helicotylenchus,

Paratylenchus Heterodera Xiphinema, Tylenchus, Criconemoides, Hemicycliophora,

Aphelenchoides, Tetylenchus, Trichodorus, Dorylaimus, Tylenchulus, Telotylenchus,

Pratylenchoides, Telotylenchoides, and Rotylenchoides. The most prominent nematode from the soil were Scutellonema with prominence value of 81.09, followed by

Meloidogyne and Rotylenchus with prominence values of 46.50 and 39.60 respectively, and from the roots, Scutellonema, Meloidogyne and Rotylenchus with prominence values of 77.15, 50.93 and 26.94, respectively. Scutellonema, Tylenchus and

Meloidogyne were the most abundant nematodes from the soil with frequency values of

97.22%, 95.83% and 86.11% respectively, while Meloidogyne, Scutellonema, and

Pratylenchus were the most abundant nematodes from the root with frequency values of

55.56%, 34.72% and 25.00% respectively,

Reaction of Jatropha curcas accessions; IARJAT2009020, IARJAT2009011,

IARJAT2009041 and IARJAT2009016 to infection with 2000 eggs of Meloidogyne incognita was evaluated under controlled environmental conditions in Samaru. There was no significant differences at P=0.05 between the four accessions with respect to

number of galls/ root at 8 weeks after inoculation and final nematode population count.

IARJAT2009016 had the highest galls/ root 10.0 followed by IARJAT2009041with 6.67 galls/ root. However, final population count had IARJAT2009011with 37.27 followed by IARJAT2009016, 28.11 and lowest population 13.50 obtained on IARJAT2009041.

Although plant parasitic nematodes were found to be associated with Jatropha curcas on the surveyed areas the pathogenicity test has shown that Meloidogyne incognita is not pathogenic on the Jatropha curcas accessions used. These accessions

(IARJAT2009020, IARJAT2009011, IARJAT2009041 and IARJAT2009016) therefore may be used to manage Meloidogyne incognita infected soils in a mixed crop combination.

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TABLE OF CONTENTS

Title Page ...... Error! Bookmark not defined.

Declaration ...... ii

Certification ...... iii

Dedication ...... iv

Acknowledgement ...... v

Abstract ...... vi

Table of Contents ...... viii

List of Tables ...... x

List of Figure ...... xii

1.0 INTRODUCTION ...... 1

1.1 Justification ...... 3

1.2 Objectives ...... 5

2.0 LITERATURE REVIEW ...... 6

2.1 Cultivation Requirements ...... 6

2.2 Botanical Description of Jatropha ...... 6

2.3 World Jatropha Production Pattern ...... 7

2.4 Uses of Jatropha ...... 7

2.5 Plant Parasitic Nematodes ...... 9

2.6 Jatropha Pests and Diseases Status ...... 12

2.7 Disease Complexes Involving Root-Knot Nematodes and Jatropha ...... 13

viii

3.0 MATERIALS AND METHODS ...... 14

3.1 Determination of Nematodes Associated with Jatropha Accessions ...... 14

3.1.1 Survey of Jatropha farms ...... 14

3.1.2 Extraction and identification of nematode genera associated with Jatropha ...... 17

3.2 Determination of Absolute Frequency, Prominence Value and Index of

Similarity ...... 17

3.3 Determination of Pathogenicity of the Root-Knot Nematode, Meloidogyne

incognita ...... 18

3.4 Data Collection ...... 20

3.5 Data Analysis ...... 21

4.0 RESULTS ...... 22

5.0 DISCUSSION ...... 47

SUMMARY, CONCLUSION AND RECOMMENDATIONS ...... 50

6.1 Summary ...... 50

6.2 Conclusion ...... 50

6.3 Recommendations ...... 51

REFERENCES ...... 52

Appendix ...... 59

LIST OF TABLES

Table 1 Top Jatropha Producers in the World 8

Table 2 Sampled Locations, soil types and their Geographical Coordinates 15

Table 3 Absolute frequency of plant parasitic nematode genera in the soil grown with Jatropha in four Local Government Areas of Kaduna 23 State, July 2013

Table 4 Absolute frequency of plant parasitic nematode genera in the root of Jatropha in four Local Government Areas of Kaduna State, July 24 2013

Table 5 Prominence value (PV) of plant parasitic nematode genera in the soil grown with Jatropha in four Local Government Areas of 26 Kaduna State, July 2013

Table 6 Prominence value (PV) of plant parasitic nematode genera in the roots of Jatropha in four Local Government Areas of Kaduna 28 State, July 2013

Table 7 Absolute frequency of plant parasitic nematode genera in the soil grown with Jatropha in four Local Government Areas of Kaduna 29 State, September 2013

Table 8 Absolute frequency of plant parasitic nematode genera in the root of Jatropha in four Local Government Areas of Kaduna State, 31 September2013

Table 9 Prominence value of plant parasitic nematode genera in the soil grown with Jatropha in four Local Government Areas of Kaduna 32 State, September 2013

Table 10 Prominence value of plant parasitic nematode genera in the root of Jatropha in four Local Government Areas of Kaduna State, 34 September 2013

Table 11 Indices of similarity for plant parasitic nematode genera in soil of Jatropha stands in six location of Kaduna State, July 2013 36

Table 12 Indices of similarity for plant-parasitic nematode genera in root of Jatropha stands in six location of Kaduna State, July 2013 37

Table 13 Indices of similarity for plant-parasitic nematode genera in soil of Jatropha stands in six locations of Kaduna State, September 2013 38

Table 14 Indices of similarity for plant-parasitic nematode genera in root of Jatropha stands in six locations of Kaduna State, September 2013 40

Table 15 Effect of Meloidogyne incognita infection on plant height of four accessions of Jatropha at Zaria, 2013 41

Table 16 Effect of Meloidogyne incognita infection on stem girth of four accessions of Jatropha 43

Table 17 Effect of Meloidogyne incognita infection on numbers of leaf counted on Jatropha plants 44

Table 18 Number of galls, population of Meloidogyne incognita and reproduction factor of inoculated Jatropha and tomato 46

LIST OF FIGURE

Figure 1 Map Showing Survey Locations in four Local Governments Areas of

Kaduna State 16

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CHAPTER ONE

1.0 INTRODUCTION

The term “Jatropha” is usually used to refer to the species Jatropha curcas, although there are approximately 170 known species of the plant (Dehgan, 1984), they include

Jatropha integerrima, Jatropha cardiophylla, Jatropha cathartica, Jatropha cinerea,

Jatropha cuneata, Jatropha podagrica and Jatropha curcas. It originated from Central

America (Jongschaap et al., 2007). It was introduced to Africa and Asia and cultivated world-wide in many parts of the tropics and subtropics where it is grown as a hedge crop and for traditional use (Heller, 1996; Kumar and Sharma, 2008). Jatropha curcas is the most common species recorded in Nigeria. Names used to describe the plant vary per region or country. It is most commonly known as “Physic nut”. In Nigeria it is known as “cinidazugu” “wuluidi” and “lapalapa” in Hausa Igbo and Yoruba languages respectively (Blench, 2007).

Jatropha curcas is a member of the family Euphorbiaceae, a large drought-resistant multipurpose shrub with several attributes and considerable potential and has evoked interest all over the tropics as a potential biofuel crop (Jones and Miller, 1991;

Openshaw, 2000). J. curcas has a straight trunk with thick branchlets. It has green leaves with a length and width of 6 cm to 15 cm. Five roots are formed from seeds: one tap root and 4 lateral roots and cuttings do not develop a taproot (Heller, 1996). The branches contain whitish latex, which causes brown stains that are difficult to remove.

The fruits is about 40 mm length and contains 3 seeds (on average), which look like black beans with similar dimensions, of about 18 mm long and 10 mm wide. The seed coats constitute about 35~40% of the total seeds (www.jartropha.org). The plant and its seeds are toxic to animals and humans and are therefore used worldwide as hedges to

protect agricultural fields. It grows on well drained soils with good aeration and is well adapted to marginal soils with low nutrient content. The best time for planting is in the warm season before or at the onset of the rains. In the former case, watering of the plants is required. The recommended spacing for plantation is 2 m to 3 m by 1.5 m to 3 m for plantations (Jones and Miller, 1992). The number of trees per hectare at planting may range from 1100 to 3300. Plant growth is dependent on soil fertility and rainfall. A poor nutrient level will lead to increased failure of seed development (Gubitz et al.,

1997). Thus, it is important to maintain soil fertility; this is contrary to statements made in some publications (Mauwa, 1995). Like all perennial plants, it displays vigorous growth at early stage of planting but this will tail off gradually towards maturity. The lifespan is more than 50 years (Openshaw, 2000). Management of Jatropha requires the addition of manure and NPK to the planting hole at 2 kg compost, 20 g urea, 120 g SSP

(single super phosphate) and 16 g MOP (muriate of potash) and urea should be applied in two splits (1 and 2 months after transplanting) at 10 g per plant (Singh et al., 1996).

Yearly top dressings of fertilizers including the seed cake should be done. The crop has been reported to have nematicidal, insecticidal, molluscidal and fungicidal properties and is expected to be less prone to damage by these pests (Anitha and Varaprasad,

2012). However, reports of pests and diseases outbreak on Jatropha started appearing with large scale cultivation in both marginal and arable lands by agro-industries (Anitha and Varaprasad, 2012).

Just like other agricultural crops, biotic factors also limit optimum production of

Jatropha. Marieke et. al., (2012), reported that over 60% of Jatropha plants fell victim to attacks by soil-borne vascular diseases. Biotic factor production constraints of Jatropha include viral, fungi and nematode infections. Nematodes are abundant in the soil, many

of which are parasites of plants including food crops and causing losses in both quantity and quality (Olabiyi et al., 2009). Plant parasitic nematodes obtain nutrients for development and reproduction from cytoplasm of the living plant cells rendering them weak and causing enormous economic losses (Stirling et al., 2002). They remain a major challenge in crop production in many developing countries including Nigeria

(Nicol et al., 2011). Some of the important plant parasitic nematodes are the: Root

Lesion Nematodes (Pratylenchus spp.), the Burrowing nematodes (Radopholus spp.) and Cyst nematodes (Heterodera and Globodera spp.). However, the most economically important nematodes, the root-knot (Meloidogyne spp.) are biotrophic which are obligate parasite that induced galls (root-knots) on the roots of their host (Van Megen et al., 2009).

1.1 Justification

Nigeria is among the countries with the largest deposits in hydrocarbons (crude oil and natural gas) and receives considerable revenue from its large multinational oil industry sectors (FGNPB, 2008). The benefits of these resources hardly trickle down to the common man, which is generally poor. Secondly, Nigeria is constantly faced with the global energy crisis characterized by oil glut with its attendant fluctuation in crude oil prices. In addition, the effect of global warming arising from emission of greenhouse gases from fossil foils poses a serious challenge both to Nigeria and the entire world.

Given these constraints, the Federal Government of Nigeria has intensified efforts aimed at diversifying the economy by looking for sources of renewable energy and alternative source of revenue (FGNPB, 2008). Jatropha curcas provides this alternative as it has been shown by other countries (Abila, 2011).

Similarly, Nigeria Government has stepped up efforts to sensitize farmers and other

Nigerians towards massive cultivation and growing of Jatropha curcas for bio-fuel production for the country to attain energy security which will at the same time, solve unemployment and poverty challenges in the country. In a bid to make Jatropha cultivation a success in Nigeria, the Renewable Energy and Energy Efficiency

Partnership (REEEP) donated a grant of € 70,000 to the Nigerian National Petroleum

Corporation (NNPC) to support a detailed feasibility study into high biofuel feedstock for production of automotive fuel (NNPC, 2005).

The adoption of Jatropha as a biofuel source holds a diversity of opportunities and potential for Nigeria economy (Abila, 2011), but the extent to which these benefits from

Jatropha production will be realized depends on many conditions which include pests and diseases. Pest and diseases have the potential to cause significant constraints on biomass production, putting the crops at risk for reductions in biomass yield and quality.

Of many pests and diseases, plant-parasitic nematodes are of great economic importance because they can directly influence plant biomass and predispose plants to attack by other soil-borne pathogens. However, few available reports on pathogenic effects of nematode on Jatropha are confusing. It was reported that pests and diseases do not pose a significant threat to Jatropha, due to the insecticidal and toxic characteristics of all parts of the plant (Heller, 1996) but despite the characteristics of the plant, incidence of pests and diseases has been widely reported under plantation monoculture, and may be of economic significance (Brittaine and Lutaladio, 2010). Since cultivation of Jatropha in Nigeria is gaining prominence, it becomes imperative to characterize the nematodes that are associated with the crop and determine the species that may be pathogenic and therefore economically important to it. This information will be vital in designing

management strategies that will ensure profitable and sustainable production of

Jatropha. It is against this background that this research was conceived.

1.2 Objectives

Study was undertaken with the following objectives:

- to determine the genera of plant parasitic nematodes associated with Jatropha

curcas in the selected Local Government Areas of Kaduna State.

- to determine the absolute frequency and prominence value of identified

nematode genera;

- to determine the pathogenicity of one of the most prominent plant parasitic

nematode genera identified on Jatropha curcas.

CHAPTER TWO

2.0 LITERATURE REVIEW

2.1 Cultivation Requirements

Jatropha is a drought-resistant, non-edible perennial. It can be grown on a marginal land, it generally grows to a height of three to five meters (Carels, 2009). Given the wide range of natural conditions it tolerates, Jatropha naturally occurs throughout the tropics and sub-tropics (Carels, 2009). The plant is well adapted to unfavorable growth conditions. In arid and semi-arid climates, it survives by shedding its leaves at the beginning of the dry season (Orwa et al., 2009).

However, with adequate water, pollinators and overall favorable growth conditions, it can thrive and produce fruits throughout the entire year (Kumar and Sharma, 2008).

Jatropha grows well in regions with an annual mean temperature between 20 to 28oC

(Orwa et al., 2009). Significantly higher water availability (minimum of 500 mm per year) is required for optimal seed production conditions because water availability is positively correlated with fruit production (Jongschaap et al., 2007).

Drained sandy or grave soils with good aeration are optimal for Jatropha as the shrub does not tolerate water logging conditions (Heller, 1996). It is well adapted to marginal soils; optimum yields are possible where sufficient amounts of nutrients are available to the plant. Jatropha is propagated by direct seeding, planting of seedling from nursery or by cuttings.

2.2 Botanical Description of Jatropha

Jatropha produces a trilocular ellipsoidal fruit. The fruits are green when unripe (Heller,

1996) and turn yellow when matured. Numerous research works on Jatropha seed and seed chemical characterization have been carried out in most Jatropha cultivation zones

(Martinez-Herrera et al., 2006; Halilu et al., 2011). Halilu et al. (2011) reported J. curcas seed weight to range from 0.28 g - 0.80 g from Nigeria and 0.45-0.72 g for seeds from Mexico (Martinez-Herrera et al., 2006). Jatropha fruit contain about 1 - 4 seeds and are black in color and about 2 cm long and 1 cm thick (Heller, 1996).

2.3 World Jatropha Production Pattern

Some Jatropha producers have reported as high as 10-15 kg seed per plant (that is 13-20 tonnes/hectare at 3.0 m x 2.5 m spacing) (Ojiako et. al., 2014). Most researchers, however, put the optimum yield at between 5-6 tonnes/ha/year under good cultivation practices (Ojiako et. al., 2014). Achten et al. (2007) put yield estimates to vary considerably, depending on the growth conditions. Achten et al. (2007) estimated output of dry seed at 1-2.5 tonnes/ha/year for land with low soil fertility and 2-5 tonnes/ha/year for land with high fertility. Oil yield in Nigeria is about 40% (Yammama, 2011).

2.4 Uses of Jatropha

Jatropha curcas is a promising species because of its useful and profitable by-products.

The cultivation of Jatropha offers the advantage of restoring soil and controlling erosion, moreover, it is suitable for intercropping, especially during the first two to five years before it starts to yield fruit (Jongschaap, 2008). Jatropha plants can serve as live fence for the protection of agricultural fields against damage by live stocks. The plant acts as cost effective bio fence compared to wire fence. It is chosen for this purpose mainly because it can easily be propagated by cuttings, densely planted for this purpose, and because the said species is not browsed by cattle. Jatropha is used as purgative/laxative, and is widely known as medicinal for treatment of a variety of ailments.

Table 1 Top Jatropha Producers in the World

S/N Country Area Planted /ha 1 China 274 559 2 India 265 422 3 Malaysia 259 906 4 Indonesia 256 545 5 Ethiopia 20 000 6 Thailand 15 680 7 Ghana 13 000 8 Burkina Faso 10 000 9 Madagascar 8 300 10 Mexico 8 040 11 Mali 8 000 12 Nigeria 7 500 13 Tanzania 6 926 14 Togo 6 000 15 Mozambique 3 585 16 Brazil 3 135 17 Uganda 3 125 18 Sri Lanka 3 000 19 Zambia 2 789 20 Senegal 2 000 21 Honduras 1 678 22 Ivory Coast 1 500 23 Kenya 1 463 24 Vietnam 1 400 25 Argentina 1 300 26 Laos 1 204 27 Dominican Rep. 1 021 28 Benin 550 29 Taiwan 536 30 Colombia 500 31 Haiti 450 32 Peru 411 33 Cambodia 395 34 El Salvador 357 35 Malawi 350 36 Paraguay 205 37 Cameroon 205 38 Guatemala 150 39 Ecuador 150 40 Caribbean 100 41 Peru 100 42 Nicaragua 63 43 Costa Rica 24

Source: Global Market Study on Jatropha (2008)

The sap flowing from the stem is used to control the bleeding of wounds. The latex of

Jatropha contains alkaloids including Jatrophine, Jatropham and curcain with anti- cancerous properties (Van den Berg et al., 1995; Thomas et al., 2008). The roots are reported as an antidote for snake-bites. The oil has a strong purgative action and is widely used to treat skin diseases and to soothe pain from rheumatism (Heller, 1996;

Marroquin et al., 1997). The oil and aqueous extract from oil has potential as an insecticide (Jain and Trivedi, 1997; Meshram et al., 1994). It has been used in the control of insect pests of cotton including cotton bollworm, and on pests of pulses, potato and corn (Kaushik and Kumar, 2004). The bark of J. curcas yields a dark blue dye which is reported to be used for coloring cloth, fishing nets and lines. The dye may be extracted from leaves and tender stems and concentrated to yellowish syrup or dried to blackish brown lumpy mass. The dye imparts to cotton different shades of tan and brown which are fairly fast. The local production of soap is one of the most economically attractive uses of Jatropha oil. The glycerin by-product of the trans- esterification process can be used to make high quality soap or it can be refined and sold at a range of prices, depending on its purity, to be used in an immense range of products, including cosmetics, toothpaste, embalming fluids, pipe joint cement, cough medicine, and tobacco (as a moistening agent). The residue after extraction (seed cake) is an excellent source of organic manure (Sharma et al., 1997). Jatropha seed cake contains curcin, a highly toxic protein similar to ricin in Castor, making it unsuitable for animal feed. Jatropha oil can be used as fuel in diesel engines directly and by blending it with methanol (Gubitz et al., 1999). Biodesel made from Jatropha oil can be used for any diesel engine without modification.

2.5 Plant Parasitic Nematodes

Over 4100 species of plant parasitic nematodes have been identified (Decraemer and 9

Hunt, 2006), and nematodes that are considered non- important are becoming important as cropping pattern changes (Nicol, 2002).

The plant parasitic nematodes of economic importance are grouped into relatively restricted specialized groups that either cause direct damage to their host or act as a virus vector. Most plant parasitic nematodes affect crops through feeding on or in plant roots (below ground), while minorities are aerial feeders (above ground) (John et al,

2013).

Among the important ones are the: Root-knot nematodes (Meloidogyne spp.) which are obligate plant parasites that have been distributed worldwide. The genus consists of 98 species (as of February 2013) and they parasitize almost every species of vascular plant

(John et al, 2013). Their common name comes from the galls (root-knots) induced by

Meloidogyne on the roots of their host plants (Moens et al., 2009).

The cyst nematodes are obligate biotrophs and are of great economic importance throughout the world. The most damaging species include soybean cyst nematodes

(Heterodera glycines), potato cyst nematodes (Globodera pallida and G. rostochiensis) and cereal cyst nematodes (Heterodera avenae and H. filipjevi). Losses caused by cereal cyst nematodes can be in excess of 90% (Nicol et al., 2011). Potato cyst nematodes have been estimated to cause losses of 9% of total potato production worldwide (Turner and Rowe, 2006). The ability of cyst nematodes to survive prolonged periods in the soil in the absence of host makes control and eradication by rotations difficult and almost impossible once established.

The root lesion nematodes (Pratylenchus spp.) There are over 60 named species which are distributed worldwide. Pratylenchus spp. rank third only to root-knot and cyst nematodes as having the greatest impacts on crop worldwide (Castillo and Volvas,

2007). The most important of these species such as P. penetrans, P. thornei,

P. neglectus, P. zeae, P. vulnus and P. coffeae. Pratylenchus are migratory, intercellular root endoparasites with a life cycle that lasts 3–8 weeks depending on the species and conditions. A reduction in root growth occurs on infection, accompanied by the formation of lesions, necrotic areas, browning and dead cell. This often followed by root rotting from secondary attack by soil fungi or bacteria. Root damage slows plant growth, increases susceptibility to water stress and causes stunting and yellowing.

Infected roots are usually discolored and stubby. Infection can occur along the entire length of the root, with damage to the epidermis, cortex and root endodermis. Their host range includes sugarcane, coffee, banana, maize, legumes, potato, many vegetables and fruit trees (Castillo and Volvas, 2007).

Dry rot nematodes (Scutellonema spp). A major economic pest of yam (Scutellonema bradys), known as the yam nematode and causal agent of yam dry rot. This nematode occurs mostly in West Africa, where yam is its principal host, but is also recorded on yam from parts of South and Central America and Asia (Bridge et al., 2005). The nematode affects all the main cultivated yam species and cultivars in West Africa

(Kwoseh, 2000; Coyne et al., 2006), Nematodes feed intracellularly in tuber tissue, rupturing cell walls and resulting in necrotic lesions.

The burrowing Nematode (Radopholus similis), of the more than 30 species in the genus Radopholus, the burrowing nematode, Radopholus similis, is the only pathogen of widespread economic importance (Duncan and Moens, 2006). Radopholus similis is a migratory endoparasitic nematode that is known to be a destructive pest of citrus crops, pepper and, most importantly, banana, on which it causes toppling disease.

Infection by R. similis results in extensive damage to root systems, which show dark lesions caused as the nematodes destructively migrate through host tissues. Tissue rot

occurs as a result of secondary bacterial and/or fungal infections, weakening the root system and leaving the plant susceptible to toppling, especially in windy conditions

(Duncan and Moens, 2006). All life stages of R. similis occur inside the roots.

Nematodes infect at or near the root tip and burrow through the root system, showing a preference for younger roots rather than those that are hardened and periodically feed on cell contents.

2.6 Jatropha Pests and Diseases Status

It was reported that pests and diseases do not pose a significant threat to Jatropha, due to the insecticidal and toxic characteristics of all parts of the plant (Heller, 1996). Despite the characteristics of the plant, incidence of pests and diseases has been widely reported under plantation monoculture, and may be of economic significance (Brittaine and

Lutaladio, 2010). There is increasing evidence that Jatropha is highly susceptible to pests and diseases which seriously hampers plant growth and seed production (Anitha and Varaprasad, 2012; Rouamba, 2011). Mekete (2010) reported that nematode species such as Xiphinema, Trichodorus, Longidorus, Helicotylenchus, Hoplolaimus,

Tylencharynchus and Pratylenchus were associated with biofuel crops including

Jatropha while Begum (1996) showed that Meloidogyne incognita is a parasite of

Jatropha podagrica. Emeh (2012) reported plant parasitic nematodes (17 genera) to be associated with Jatropha curcas on two fields located on the Institute for Agricultural

Research, Ahmadu Bello University, Zaria, and concluded that Jatropha curcas is a host of many plant parasitic nematodes. There have been reports on collar rot disease

(caused by Macropphomina phaseolina or Rhizoctonia bataticola) at juvenile stages or by water-logging at adult stages, leaf spots disease, root rot disease (caused by

Fusarium moniliforme) and damping off (caused by Phytophtora spp.) (Sharma and

Sarraf, 2007). Dieback/stem-rot was reported on Jatropha seedling by Zarafi and

Abdulkadir (2013).

2.7 Disease Complexes Involving Root-Knot Nematodes and Jatropha

A number of disease complexes involving many crops in which root-knot nematodes are recorded interacting with fungal pathogens have been reported (McGaweley, 2001).

Cultivars of Jatropha with known resistance to fungal pathogens becomes susceptible, either completely or partially, when such cultivars are simultaneously parasitized by plant parasitic nematodes (McGaweley, 2001). In certain situations, the nematode has been found to be responsible for breaking disease resistance to Fusarium wilt

(McGaweley, 2001). Studies on Jatropha have shown that new diseases such as viral diseases causing floral deformity are being discovered to have serious damage on the plant (Gour, 2004). Plant parasitic nematodes do not only attack the plant directly but also predispose them to attack by other soil-borne pathogens (Mekete, 2010).

CHAPTER THREE

3.0 MATERIALS AND METHODS

3.1 Determination of Nematodes Associated with Jatropha Accessions

3.1.1 Survey of Jatropha farms

Six locations distributed over four Local Government Areas of Kaduna State were surveyed to determine the different plant parasitic nematodes associated with Jatropha

(Table 2 and Fig. 1). Coordinate of surveyed areas were collected using phone software called „Open Data Kit‟. Plant samples were taken from two fields in each location. In the Zaria and Basawa orchards, soil samples were collected from Jatropha orchards using the system random sampling method. Soil samples were collected from 6 plant stands spaced 8 m apart to make a sub-sample. Samples were taken from each plant base at a distance of 25 cm from the plant. Soil samples at a depth of 25 cm were removed from the root zone using soil auger and hoe. Ten (10) sub-samples were bulk together to make a bulk sample and a total number of 6 bulk samples were taken from each field. In the farmers‟ fields, soil samples were collected from 4 plants stands to make a sub-sample, 6 sub-samples were bulk together to make a bulk sample. Root samples were also collected with scissors from each Jatropha stand where soil was collected at a depth of 25 cm. The root tips were slightly exposed, cut with scissors and covered back with soil. Samples were bagged in a polythene bag, labeled for identification, sealed and taken to the Nematology Laboratory, Department of Crop

Protection, Ahmadu Bello University, Zaria and stored in the Cold Room before extraction. Samples were collected at the peak of rainy season (July) and towards the end of the rainy season (September) to check the differences in nematode population.

Table 2 Sampled Locations, soil types and their Geographical Coordinates

S/NO Location Local Soil Type Coordinates Government Area Latitude Longitude 1 Jatropha Plantation of the Sabon Gari Sandy loam 11° 9' 59.8" 7° 37' 59.5" Institute for Agricultural Research, (IAR), Samaru, Ahmadu Bello University, Zaria, Zaria. 2 Jatropha Plantation of Sabon Gari Sandy loam 11o 9' 8.2" 7o 41' 08.1" National Research Institute for Chemical Technology (NARICT), Basawa. 3 Farmer‟s field in Kurmi Sabon Gari Sandy loam 11o 0' 4.7" 7o 37' 49.6" Bomo

4 Farmer‟s field in Hunkuyi Kudan Sandy loam 11o 15' 0" 7o 38' 51.9"

5 Farmer‟s field in Giwa Giwa Loamy 11o 16' 1" 7o 25' 01.9"

6 Farmer‟s field in Unguwar Zaria Sandy loam 11o 4' 45 " 7o 40' 57" Dankali

Source: eHealth Africa, GIS Dept, Kano State, Nigeria

Figure 1 Surveyed Locations in four Local Government Areas of Kaduna State

3.1.2 Extraction and identification of nematode genera associated with Jatropha

Nematodes were extracted from 72 soil samples within two weeks of collection using

Cobb‟s (1918) modified Sieving and Decanting technique. Nematodes from the roots were extracted using Baermann pan method. The final suspensions containing the nematodes in both samples were viewed under the light microscope and nematodes identification chart described by Mai and Mullin, (1996) was used to identify the nematodes.

3.2 Determination of Absolute Frequency, Prominence Value and Index of

Similarity

Data on nematode numbers was analyzed using absolute frequency (AF), prominence value (PV) (Norton, 1989) and Index of Similarity (IS) (Bray and Curtis, 1957)

Absolute frequency is an independent calculation that denotes how often a genus occurs among the samples examined.

Thus Absolute Frequency (AF) = 100 where g = number of samples containing a genus n = total number of samples collected.

Prominence Value is the density of a population multiplied by the square root of the absolute frequency.

Prominence Value (PV) = d where d = nematode density,

AF = absolute frequency

Index of Similarity is the measure of how close two sampled locations are to one another in terms of nematode genera recorded.

Index of Similarity (IS) = 2c ∕ (a + b) where c = number of genera two locations have in common, a and b = the number of genera in zones a and b respectively

3.3 Determination of Pathogenicity of the Root-Knot Nematode, Meloidogyne incognita

Soil Sterilization

Thoroughly mixed loam, sand and manure in the ratio 2:1:1 was heat-sterilized by loading moistened soil in a longitudinally cut drum on burning fire for 90 mins with regularly turning at every 30 mins to ensure even distribution of heat. Sterilized soil was dried and utilized one week later.

Tomato plants infected with root-knot nematodes were collected from farmer‟s plots.

These were examined for females and single egg masses under the dissecting microscope after thorough washing under a stream of tap water. Females of

Meloidogyne spp. were distinguished using perineal patterns (Eisenback et al., 1981).

Egg masses of identified M. incognita females were used to inoculate by placing the egg masses close to growing tip of roots of susceptible tomato variety Roma-VF. After eight weeks of inoculation, females were isolated from these plants and perineal patterns were made to ascertain purity of the culture (M. incognita). Subsequently, pure cultures were multiplied in pots (15 cm diameter × 22 cm height) with three plants grown in each.

These were used for subsequent experiment.

Four Jatropha curcas accessions seeds namely: IARJAT2009020, IARJAT2009016,

IARJAT2009041 and IARJAT2009012 obtained from the Legumes and Oilseeds

Research Programe of the Institute for Agricultural Research (Samaru), Zaria and

Roma-VF obtained from Premier Seeds Ltd. Zaria, were raised in separate nursery trays

100 cm x 50 cm quarter filled with heat sterilized soil. Jatropha and tomato seeds at distance of 3cm were finely spread along furrows before it was lightly covered and mulched with dry grass. The tomato and Jatropha seeds were watered at 24 hours interval.

Four (4) kilograms of sterilized soil was weighed into each plastic pot 15 cm diameter ×

22 cm height, perforated at base for drainage. Two weeks old Jatropha and one week old Roma-VF tomato seedlings were transplanted at a rate of 2 seedlings per pot. Two seedlings were thinned to only one per pot five days after transplanting.

Un-inoculated Jatropha curcas accessions were the control treatments while Roma VF served as the standard check. The experiment was replicated six times. Sixty (60) pots were arranged in a Completely Randomized Design (CRD) in the screen-house.

Meloidogyne incognita eggs obtained from the pure culture were used to inoculate each

Jatropha and tomato seedlingsss. Meloidogyne incognita eggs were extracted from pure culture using Sodium Hypochlorite (NaOCl) as described by Hussey and Baker (1973).

Galled roots from 42 tomato plants were washed of adhering soils, mopped dry with clean soft tissue paper and cut into 1-2 cm pieces. Root pieces were placed in 1000 ml conical flask, 0.52 %NaOCl as diluent was added at a ratio 1:3 of water. The flask was tightly corked, shaken vigorously for 2 mins to dissolve gelatinous matrix enclosing the eggs. Suspension containing the eggs and juveniles was poured over 0.075 mm size mesh nested upon 0.045 mm sieve. The sieve (0.045 mm) containing eggs/ juveniles

(J2) was placed under a gentle stream of water to rinse off residual NaOCl. Rinsed egg and J2 larvae in the mesh were washed with water into a wash bottle and adjusted to a known volume (360 ml). Using dissecting microscope, counts of eggs and J2 larvae per

10 ml aliquot was done using Doncaster counting dish. Inoculation of seedlings was done 2 weeks after transplanting (WAT) by making Shallow trench around base of each transplant using clean spatula. Approximately 2000 egg suspension was poured into the trench, cover with soil and water lightly (Iheukwumere et al., 1995).

Data on plant height, stem girth, number of leaves at 14 days interval for eight weeks after inoculation (WAI) were collected. Tomato variety, Roma VF was used as standard check. Eight weeks after inoculation, soil in the pots were loosened manually, roots and soil were taken to the laboratory for assessment.

3.4 Data Collection

Roots of Jatropha and tomato plants were washed under a gentle stream of tap water, wrapped with soft tissue paper, labeled and arranged on a table, scored for galling using the 0-5 rating scale described by Taylor and Sasser (1978) presented below:

Score Gall Rating

0 No gall on root

1 1 – 2 galls on root

2 3 – 10 galls on root

3 11 – 30 galls on root

4 31 – 100 galls on root

5 > 100 galls on root

Direct counting technique was employed to get number of galls and matured egg masses per gram of root (Daykin and Hussey, 1985). Root of each plant was cut into pieces of about 1-2 cm long and thoroughly mixed. Using a Mettler weighing balance, 2 g of root was weighed from thoroughly mixed cut roots. Root pieces were spread evenly in a grid

Petri-dish. Direct counting of number of galls on these roots was done and recorded.

Number of matured egg masses was also counted and recorded. Results obtained were then converted to per gram of root weight.

Nematode extraction from the soil was carried out using Sieving and Decanting method described by Cobb (1918), to determine the reproduction factor (RF) which is the ratio of final population to initial population (Oostenbrink, 1966).

RF = Final Population

Initial Population

3.5 Data Analysis

Data collected were subjected to Statistical Analysis of Variance (ANOVA) using the

SAS Software Version 9.4 (2013). Means were separated using the Duncan Multiple

Range Test (DMRT) at 5% level of probability (P=0.05).

CHAPTER FOUR

4.0 RESULTS

The absolute frequency of nematodes genera in soils grown with Jatropha curcas accessions is presented in Table 3. In July 2013, twenty-four (24) genera of plant parasitic nematodes were recorded from the six locations. Nematodes recorded were

Scutellonema, Hoplolaimus, Pratylenchus, Aphelenchus, Meloidogyne,

Tylenchoryhnchus, Rotylenchus, Longidorus, Helicotylenchus, Paratylenchus,

Heterodera , Xiphinema, Tylenchus, Criconemoides, Hemicycliophora, Aphelenchoides,

Tetylenchus, Trichodorus, Dorylaimus, Tylenchulus, Telotylenchus, Pratylenchoides,

Telotylenchoides and Rotylenchoides. At all the locations, Scutellonema had the highest frequency value (97.22), followed by Tylenchus and Meloidogyne with frequency values of 95.83 and 86.11 respectively. Dorylaimus and Rotylenchoides were the least nematode genera recorded with frequency value of 1.39 each. This is because

Dorylaimus and Rotylenchoides were only recorded in one location, (Basawa) out of the six locations surveyed. The locations surveyed i.e. Basawa, IAR Samaru, Giwa,

Unguwar Dankali, Kurmi Bomo and Hunkuyi recorded 22, 19, 18, 19, 19 and 17 genera of nematodes respectively. Telotylenchus, Pratylenchoides, Telotylenchoides and

Tylenchulus were not recorded in five (5) locations namely IAR Samaru, Giwa,

Unguwar Dankali, Kurmi Bomo and Hunkuyi while the populations encountered at

Basawa were low. Dorylaimus, Telotylenchus, Pratylenchoides, Telotylenchoides and

Rotylenchoides were not common to all the locations but encountered at Kurmi Bomo,

Basawa and IAR Samaru.

Absolute frequency of nematodes genera in the root of Jatropha curcas accessions in

July 2013 is shown in Table 4. Eleven (11) genera of plant parasitic nematodes were recorded in the root of Jatropha obtained from the six locations. Scutellonema,

Table 3 Absolute frequency of plant parasitic nematode genera in the soil grown with Jatropha in four Local Government Areas of Kaduna State, July 2013

Absolute Frequency (%) Nematode Genera Locations

Basawa IAR Samaru Giwa Unguwar Dankali Kurmi Bomo Hunkuyi All Locations

(n=12) (n=12) (n=12) (n=12) (n=12) (n=12) (n=72) Scutellonema 100.00 100.00 100.00 100.00 91.67 91.67 97.22 Hoplolaimus 75.00 50.00 83.33 83.33 91.67 83.33 77.78 Pratylenchus 66.67 91.67 58.33 83.33 66.67 58.33 70.83 Aphelenchus 91.67 91.67 83.33 58.33 91.67 83.33 83.33 Meloidogyne 100.00 75.00 83.33 83.33 91.67 83.33 86.11 Tylenchoryhnchus 16.67 25.00 16.67 41.67 25.00 8.33 22.22 Rotylenchus 91.67 66.67 83.33 83.33 91.67 83.33 83.33 Longidorus 75.00 75.00 91.67 75.00 91.67 91.67 83.33 Helicotylenchus 83.33 75.00 25.00 66.67 66.67 41.67 59.72 Paratylenchus 58.33 91.67 50.00 33.33 33.33 58.33 54.17 Heterodera 25.00 0.00 8.33 16.67 16.67 8.33 12.50 Xiphinema 83.33 100.00 83.33 83.33 91.67 58.33 83.33 Tylenchus 91.67 100.00 91.67 91.67 100.00 100.00 95.83 Criconemoides 8.33 50.00 41.67 58.33 50.00 33.33 40.28 Hemicycliophora 50.00 41.67 58.33 41.67 33.33 50.00 45.83 Aphelenchoides 83.33 50.00 75.00 75.00 75.00 50.00 68.06 Tetylenchus 25.00 16.67 8.33 16.67 16.67 16.67 16.67 Trichodorus 0.00 33.33 0.00 16.67 25.00 0.00 12.50 Dorylaimus 0.00 0.00 0.00 0.00 8.33 0.00 1.39 Tylenchulus 58.33 25.00 66.67 33.33 0.00 0.00 30.56 Telotylenchus 25.00 0.00 0.00 0.00 0.00 0.00 4.17 Pratylenchoides 33.33 0.00 0.00 0.00 0.00 0.00 5.56 Telotylenchoides 16.67 16.67 0.00 0.00 0.00 0.00 5.56 Rotylenchoides 8.33 0.00 0.00 0.00 0.00 0.00 1.39

Table 4 Absolute frequency of plant parasitic nematode genera in the root of Jatropha in four Local Government Areas of Kaduna State, July 2013

Absolute Frequency (%) Locations

Nematode Genera Basawa IAR Samaru Giwa Unguwar Dankali Kurmi Bomo Hunkuyi All Locations

(n=12) (n=12) (n=12) (n=12) (n=12) (n=12) (n=72)

Scutellonema 25.00 50.00 66.67 58.33 8.33 0.00 34.72

Hoplolaimus 8.33 16.67 33.33 33.33 8.33 0.00 16.67

Pratylenchus 66.67 16.67 16.67 41.67 8.33 0.00 25.00

Meloidogyne 66.67 75.00 58.33 75.00 50.00 8.33 55.56

Rotylenchus 16.67 33.33 0.00 16.67 0.00 0.00 11.11

Longidorus 16.67 25.00 25.00 25.00 0.00 0.00 15.28

Xiphinema 0.00 33.33 25.00 33.33 0.00 0.00 15.28

Aphelenchoides 0.00 0.00 0.00 8.33 0.00 0.00 1.39

Heterodera 8.33 0.00 0.00 0.00 0.00 0.00 1.39

Helicotylenchus 8.33 0.00 0.00 0.00 0.00 0.00 1.39

Paratylenchus 8.33 0.00 0.00 0.00 0.00 0.00 1.39

Hoplolaimus, Pratylenchus, Meloidogyne, Rotylenchus, Longidorus, Helicotylenchus,

Paratylenchus Heterodera, Xiphinema and Aphelenchoides were the genera recorded.

Meloidogyne had the highest frequency value, 55.56 in all the locations, followed by

Scutellonema, and Pratylenchus with frequency values of 34.72 and 25.00 respectively.

Aphelenchoides, Heterodera, Helicotylenchus and Paratylenchus were the least nematode genera encountered with frequency value of 1.39 each. The locations Basawa,

IAR Samaru, Giwa, Unguwar Dankali, Kurmi Bomo and Hunkuyi recorded 9, 7, 6, 8, 4 and 1 genera of nematodes respectively. Meloidogyne was the only nematode genus obtained at Hunkuyi while Heterodera, Helicotylenchus and Paratylenchus were recorded in Basawa. Unguwar Dankali was the only location where Aphelenchoides was recorded. Scutellonema, Hoplolaimus, Pratylenchus and Meloidogyne were common to

Basawa, IAR Samaru, Giwa, Unguwar Dankali and Kurmi Bomo.

Prominence value of nematodes genera encountered in soil grown with Jatropha curcas accession in July, 2013 are presented in Table 5. Twenty-four (24) genera of plant parasitic nematodes were recorded. Scutellonema, Hoplolaimus, Pratylenchus,

Aphelenchus, Meloidogyne, Tylenchoryhnchus, Rotylenchus, Longidorus,

Helicotylenchus, Paratylenchus Heterodera Xiphinema, Tylenchus, Criconemoides,

Hemicycliophora, Aphelenchoides, Tetylenchus, Trichodorus, Dorylaimus, Tylenchulus,

Telotylenchus, Pratylenchoides, Telotylenchoides, and Rotylenchoides were the nematode recorded. Out of the 24 genera, Scutellonema, Meloidogyne and Rotylenchus were more prominent with prominence values of 81.09, 46.50 and 39.60 respectively.

Dorylaimus was the least prominent genera (0.63) and was recorded only in Kurmi

Bomo. Locations such as Basawa, IAR Samaru, Giwa, Unguwar Dankali, Kurmi Bomo and Hunkuyi recorded 22, 19, 18, 19, 19 and 17 genera of nematodes respectively.

Telotylenchus, Pratylenchoides, Telotylenchoides and Rotylenchoides were not recorded

Table 5 Prominence value (PV) of plant parasitic nematode genera in the soil grown with Jatropha in four Local Government Areas of Kaduna State, July 2013

Prominence value Nematode Genera Locations

Basawa IAR Samaru Giwa Unguwar Dankali Kurmi Bomo Hunkuyi All Locations (n=12) (n=12) (n=12) (n=12) (n=12) (n=12) (n=72) Scutellonema 105.82 73.04 52.82 86.88 90.43 77.89 81.09 Hoplolaimus 41.67 20.07 24.34 51.06 37.75 33.47 35.11 Pratylenchus 25.31 30.75 21.75 38.89 28.71 36.73 30.44 Aphelenchus 36.90 23.67 27.63 29.46 32.32 23.31 28.84 Meloidogyne 49.46 54.23 39.38 52.28 43.79 40.41 46.50 Tylenchoryhnchus 6.12 12.22 3.95 9.81 5.56 3.85 7.23 Rotylenchus 44.59 29.48 32.44 56.42 54.80 18.14 39.60 Longidorus 27.65 18.99 21.18 18.41 15.43 12.30 18.84 Helicotylenchus 27.08 22.58 7.56 45.52 21.02 22.38 25.20 Paratylenchus 20.66 14.22 16.34 9.33 9.91 15.57 14.48 Heterodera 3.11 0.00 3.85 2.45 2.31 1.73 2.41 Xiphinema 30.49 22.94 16.01 16.61 14.16 12.29 18.90 Tylenchus 28.66 35.00 31.80 23.38 27.83 24.78 28.58 Criconemoides 1.54 8.49 6.37 7.71 5.11 5.39 6.08 Hemicycliophora 11.71 11.88 19.79 9.73 9.33 7.86 11.89 Aphelenchoides 20.94 8.01 12.38 13.34 13.73 12.18 13.65 Tetylenchus 5.00 3.67 1.54 2.59 6.26 3.95 3.95 Trichodorus 0.00 4.62 0.00 3.40 3.33 0.00 2.70 Dorylaimus 0.00 0.00 0.00 0.00 1.54 0.00 0.63 Tylenchulus 12.07 4.44 8.57 4.81 0.00 0.00 6.40 Telotylenchus 5.22 0.00 0.00 0.00 0.00 0.00 2.13 Pratylenchoides 7.31 0.00 0.00 0.00 0.00 0.00 2.99 Telotylenchoides 6.12 3.54 0.00 0.00 0.00 0.00 2.79 Rotylenchoides 3.66 0.00 0.00 0.00 0.00 0.00 1.49 26

in Giwa, Unguwar Dankali, Kurmi Bomo, Hunkuyi and IAR Samaru but

Telotylenchoides was recorded in IAR Samaru. Nematodes, Trichodorus, Dorylaimus,

Telotylenchus, Pratylenchoides, Telotylenchoides and Rotylenchoides, that were absent in most of the locations have low prominent values that ranges between 0.63 and 2.99.

Seventeen (17) nematode genera were common to all the locations.

Prominence value of nematodes genera in the root of Jatropha curcas accession in July

2013 is shown in Table 6. Eleven (11) genera of plant parasitic nematodes recorded are

Scutellonema, Hoplolaimus, Pratylenchus, Meloidogyne, Rotylenchus, Longidorus,

Helicotylenchus, Paratylenchus Heterodera, Xiphinema and Aphelenchoides.

Scutellonema, Meloidogyne, Rotylenchus and Pratylenchus were more prominent with prominence values of 77.15, 50.93, 26.94 and 26.57 respectively. Aphelenchoides and

Paratylenchus were the least prominent nematodes genera with a prominence value of

3.14 each and were recorded in Unguwar Dankali and Basawa respectively. Basawa,

IAR Samaru, Giwa, Unguwar Dankali, Kurmi Bomo and Hunkuyi had 9, 7, 6, 8, 4 and

1 genera of nematodes respectively. Genus Meloidogyne was the only nematode recorded at Hunkuyi. Heterodera, Helicotylenchus and Paratylenchus were absent in 5 locations but found in Basawa. Aphelenchoides was recorded only in Unguwar Dankali while Scutellonema, Hoplolaimus, Pratylenchus and Meloidogyne were common to all the locations except Hunkuyi that has only one (1) nematode genera.

Absolute frequency of nematodes genera associated with Jatropha curcas accession grown in the soil in September, 2013 is shown in Table 7. Twenty (20) genera of plant parasitic nematodes were recorded. The nematodes recorded were Scutellonema,

Hoplolaimus, Pratylenchus, Aphelenchus, Meloidogyne, Tylenchoryhnchus,

Rotylenchus, Longidorus, Helicotylenchus, Paratylenchus Heterodera Xiphinema,

Table 6 Prominence value (PV) of plant parasitic nematode genera in the roots of Jatropha in four Local Government Areas of Kaduna State, July 2013

Prominence value Locations

Nematode Genera Basawa IAR Samaru Giwa Unguwar Dankali Kurmi Bomo Hunkuyi All Locations

(n=12) (n=12) (n=12) (n=12) (n=12) (n=12) (n=72)

Scutellonema 65.00 118.64 65.66 123.66 28.87 0.00 77.15

Hoplolaimus 9.62 13.61 22.61 26.46 8.66 0.00 15.99

Pratylenchus 42.87 25.86 12.25 41.31 8.66 0.00 26.57

Meloidogyne 68.04 61.90 44.37 58.38 44.39 9.62 50.93

Rotylenchus 20.41 48.11 0.00 43.55 0.00 0.00 26.94

Longidorus 12.93 15.00 15.56 16.11 0.00 0.00 12.20

Xiphinema 0.00 17.80 22.22 22.61 0.00 0.00 14.69

Aphelenchoides 0.00 0.00 0.00 7.70 0.00 0.00 3.14 Heterodera 9.62 0.00 0.00 0.00 0.00 0.00 3.93

Helicotylenchus 9.62 0.00 0.00 0.00 0.00 0.00 3.93

Paratylenchus 7.70 0.00 0.00 0.00 0.00 0.00 3.14

Table 7 Absolute frequency of plant parasitic nematode genera in the soil grown with Jatropha in four Local Government Areas of Kaduna State, September 2013

Absolute Frequency (%) Nematode Genera Locations

Basawa IAR Samaru Giwa Unguwar Dankali Kurmi Bomo Hunkuyi All Locations

(n=12) (n=12) (n=12) (n=12) (n=12) (n=12) (n=72) Scutellonema 91.67 100.00 66.67 100.00 100.00 83.33 90.28 Hoplolaimus 100.00 91.67 83.33 91.67 100.00 66.67 88.89 Pratylenchus 83.33 66.67 75.00 91.67 33.33 41.67 65.28 Aphelenchus 58.33 91.67 66.67 83.33 91.67 50.00 73.61 Meloidogyne 91.67 83.33 83.33 83.33 91.67 83.33 86.11 Tylenchoryhnchus 16.67 33.33 8.33 16.67 16.67 8.33 16.67

Rotylenchus 75.00 91.67 41.67 58.33 100.00 58.33 70.83 Longidorus 83.33 75.00 66.67 91.67 83.33 66.67 77.78 Helicotylenchus 83.33 66.67 50.00 58.33 50.00 41.67 58.33 Paratylenchus 16.67 58.33 41.67 75.00 16.67 66.67 45.83 Heterodera 8.33 16.67 8.33 8.33 33.33 8.33 13.89 Xiphinema 91.67 75.00 83.33 66.67 91.67 58.33 77.78 Tylenchus 100.00 100.00 83.33 100.00 100.00 100.00 97.22 Criconemoides 58.33 25.00 16.67 33.33 41.67 16.67 31.94 Hemicycliophora 33.33 75.00 58.33 66.67 41.67 25.00 50.00 Aphelenchoides 91.67 58.33 50.00 33.33 66.67 41.67 56.94 Tetylenchus 0.00 41.67 25.00 25.00 25.00 25.00 23.61 Trichodorus 8.33 0.00 0.00 0.00 8.33 0.00 2.78 Dorylaimus 8.33 0.00 0.00 0.00 0.00 0.00 1.39 Tylenchulus 0.00 8.33 75.00 0.00 33.33 25.00 23.61 Telotylenchus 0.00 0.00 0.00 0.00 0.00 0.00 0.00 Pratylenchoides 0.00 0.00 0.00 0.00 0.00 0.00 0.00 Telotylenchoides 0.00 0.00 0.00 0.00 0.00 0.00 0.00 Rotylenchoides 0.00 0.00 0.00 0.00 0.00 0.00 0.00

Tylenchus, Criconemoides, Hemicycliophora, Aphelenchoides, Tetylenchus,

Trichodorus, Dorylaimus and Tylenchulus. In all the locations, Tylenchus, Scutellonema,

Hoplolaimus and Meloidogyne were the most frequently occurring nematode genera with frequency values of 97.22, 90.28, 88.89 and 86.11 respectively. Genus Dorylaimus has the least frequency value (1.39), and it was recorded in one location (Basawa) out of the six locations surveyed. The locations Basawa, IAR Samaru, Giwa, Unguwar

Dankali, Kurmi Bomo and Hunkuyi had 18, 18, 18, 17, 19 and 18 genera of nematodes respectively recorded. Trichodorus was recorded at Basawa and Kurmi Bomo while

Dorylaimus was recorded only at Basawa. Out of the 20 genera of nematodes recorded, only Dorylaimus and Trichodorus were not common to all the locations.

The result of the absolute frequency of nematodes genera in the root of Jatropha curcas accession in September 2013 is presented in Table 8. Six (6) genera of plant parasitic nematodes were recorded in the root of Jatropha curcas. Scutellonema, Hoplolaimus,

Pratylenchus, Rotylenchus, Longidorus, and Paratylenchus were the genera recorded.

Scutellonema had the highest frequency value of 13.89, followed by Hoplolaimus,

Pratylenchus, Rotylenchus and Paratylenchus with frequency values of 4.17 each.

Longidorus was the least nematode genera recorded, with a frequency value of 1.39.

The locations, Basawa, IAR Samaru, Unguwar Dankali and Kurmi Bomo recorded 3, 5,

3 and 2 genera of nematodes respectively. No nematode was recorded in Giwa and

Hunkuyi. Longidorus was the only nematode genus recorded in Hunkuyi. Scutellonema was the only nematode common to the four locations where nematodes were recorded.

The prominence value of nematodes genera in the root of Jatropha curcas accession in

September, 2013 is shown in Table 9. Twenty (20) genera of plant parasitic nematodes were recorded. Scutellonema, Hoplolaimus, Pratylenchus, Aphelenchus, Meloidogyne,

Tylenchoryhnchus, Rotylenchus, Longidorus, Helicotylenchus, Paratylenchus

Table 8 Absolute frequency of plant parasitic nematode genera in the root of Jatropha in four Local Government Areas of Kaduna State, September 2013

Absolute Frequency (%) Locations

Nematode Genera IAR Unguwar Kurmi All Basawa Giwa Hunkuyi Samaru Dankali Bomo Locations

(n=12) (n=12) (n=12) (n=12) (n=12) (n=12) (n=72)

Scutellonema 16.67 25.00 0.00 25.00 16.67 0.00 13.89

Hoplolaimus 0.00 8.33 0.00 16.67 0.00 0.00 4.17

Pratylenchus 8.33 16.67 0.00 0.00 0.00 0.00 4.17

Meloidogyne 0.00 0.00 0.00 0.00 0.00 0.00 0.00

Rotylenchus 0.00 16.67 0.00 8.33 0.00 0.00 4.17

Longidorus 8.33 0.00 0.00 0.00 0.00 0.00 1.39

Xiphinema 0.00 0.00 0.00 0.00 0.00 0.00 0.00

Aphelenchoides 0.00 0.00 0.00 0.00 0.00 0.00 0.00

Heterodera 0.00 0.00 0.00 0.00 0.00 0.00 0.00

Helicotylenchus 0.00 0.00 0.00 0.00 0.00 0.00 0.00

Paratylenchus 0.00 16.67 0.00 0.00 8.33 0.00 4.17

Table 9 Prominence value of plant parasitic nematode genera in the soil grown with Jatropha in four Local Government Areas of Kaduna State, September 2013

Prominence value Nematode Genera Locations Basawa IAR Giwa Unguwar Kurmi Bomo Hunkuyi All Locations (n=12) Samaru (n=12) Dankali (n=12) (n=12) (n=72) (n=12) (n=12) Scutellonema 155.65 276.04 29.88 73.21 137.66 40.50 122.31 Hoplolaimus 149.60 28.46 12.54 23.29 59.71 25.73 51.50 Pratylenchus 75.71 22.05 15.85 16.77 31.18 6.31 28.67 Aphelenchus 40.59 28.66 10.63 10.22 45.32 15.32 25.39 Meloidogyne 51.45 38.89 39.40 51.90 41.76 40.94 44.08 Tylenchoryhnchus 5.44 9.24 3.85 3.95 9.53 3.46 6.19 Rotylenchus 90.55 82.30 14.11 13.95 90.88 18.90 55.90 Longidorus 14.55 10.01 13.74 10.91 25.62 10.21 14.24 Helicotylenchus 142.89 22.39 5.75 10.33 28.91 45.01 45.86 Paratylenchus 6.12 7.27 6.45 10.71 14.56 11.50 9.07 Heterodera 1.54 2.45 1.73 1.73 5.48 1.73 2.73 Xiphinema 20.89 9.49 12.98 7.21 14.39 7.69 12.40 Tylenchus 35.33 23.00 12.64 30.67 32.39 15.11 25.02 Criconemoides 9.67 2.89 2.45 3.75 5.68 2.31 4.90 Hemicycliophora 16.07 9.37 8.20 7.08 6.11 4.11 8.35 Aphelenchoides 24.31 6.47 6.91 5.29 19.60 4.43 12.15 Tetylenchus 0.00 3.96 3.11 5.11 3.11 3.79 3.47 Trichodorus 3.85 0.00 0.00 0.00 1.92 0.00 1.67 Dorylaimus 1.54 0.00 0.00 0.00 0.00 0.00 0.63 Tylenchulus 0.00 1.92 9.00 0.00 5.10 5.11 4.75 Telotylenchus 0.00 0.00 0.00 0.00 0.00 0.00 0.00 Pratylenchoides 0.00 0.00 0.00 0.00 0.00 0.00 0.00 Telotylenchoides 0.00 0.00 0.00 0.00 0.00 0.00 0.00 Rotylenchoides 0.00 0.00 0.00 0.00 0.00 0.00 0.00

Heterodera, Xiphinema, Tylenchus, Criconemoides, Hemicycliophora, Aphelenchoides,

Tetylenchus, Trichodorus, Dorylaimus, and Tylenchulus, were the nematode recorded.

Out of the 20 genera, Scutellonema was the most prominent (122.31) nematode genera recorded in all the locations. It was followed by Rotylenchus, Hoplolaimus,

Helicotylenchus and Meloidogyne with prominence values of 55.90, 51.50, 45.86 and

44.08 respectively. Dorylaimus was the least prominent nematode genera (0.63) and it was recorded only in Basawa. Eighteen (18), 18, 18, 17,19 and 18 nematode genera were recorded in Basawa, IAR Samaru, Giwa, Unguwar Dankali, Kurmi Bomo and

Hunkuyi respectively. Telotylenchus, Pratylenchoides, Telotylenchoides and

Rotylenchoides were not recorded in all the locations. Scutellonema, Hoplolaimus,

Pratylenchus, Aphelenchus, Meloidogyne, Tylenchoryhnchus, Rotylenchus, Longidorus,

Helicotylenchus, Paratylenchus Heterodera Xiphinema, Tylenchus, Criconemoides,

Hemicycliophora and Aphelenchoides were common to all the locations.

The prominence values of nematodes genera in the root of Jatropha curcas accession in

September 2013 is shown in Table 10. Six (6) genera of plant parasitic nematodes were recorded in the root of Jatropha. Scutellonema, Hoplolaimus, Pratylenchus,

Rotylenchus, Longidorus, and Paratylenchus were the genera recorded. Scutellonema was more prominent (21.49) out of the 6 genera recorded. Paratylenchus, Hoplolaimus,

Pratylenchus and Rotylenchus were also prominent with prominence values of 6.35,

6.12, 5.90 and 5.67 respectively. Longidorus was the least prominent nematode genera recorded with a prominence value of 3.54. Basawa, IAR Samaru, Unguwar Dankali and

Kurmi Bomo had 3, 5, 3, and 2 genera of nematodes respectively recorded while no nematode was encountered at Giwa and Hunkuyi. Xiphinema, Aphelenchoides,

Heterodera and Helicotylenchus were not recorded in any of the locations while

Longidorous was recorded only in Basawa. Hoplolaimus and Rotylenchus were

Table 10 Prominence value of plant parasitic nematode genera in the root of Jatropha in four Local Government Areas of Kaduna State, September 2013

Prominence value

Locations

Nematode Genera Kurmi All Basawa IAR Samaru Giwa Unguwar Dankali Bomo Hunkuyi Locations

(n=12) (n=12) (n=12) (n=12) (n=12) (n=12) (n=72)

Scutellonema 12.25 27.22 0.00 42.22 20.41 0.00 21.49

Hoplolaimus 0.00 7.70 0.00 12.93 0.00 0.00 6.12

Pratylenchus 8.66 11.57 0.00 0.00 0.00 0.00 5.90

Meloidogyne 0.00 0.00 0.00 0.00 0.00 0.00 0.00

Rotylenchus 0.00 11.57 0.00 7.70 0.00 0.00 5.67

Longidorus 8.66 0.00 0.00 0.00 0.00 0.00 3.54

Xiphinema 0.00 0.00 0.00 0.00 0.00 0.00 0.00

Aphelenchoides 0.00 0.00 0.00 0.00 0.00 0.00 0.00

Heterodera 0.00 0.00 0.00 0.00 0.00 0.00 0.00

Helicotylenchus 0.00 0.00 0.00 0.00 0.00 0.00 0.00

Paratylenchus 0.00 13.61 0.00 0.00 7.70 0.00 6.35

recorded in IAR Samaru and Unguwar Dankali. Scutellonema was the only nematode genus common to all locations (Basawa, IAR Samaru, Unguwar Dankali and Kurmi

Bomo).

For July 2013, the index of similarity between the locations (Basawa, IAR Samaru,

Giwa, UngwaDankali, Kurmi Bomo and Hunkuyi) and number of nematodes they have in common in soil is presented in Table 11. All locations were similar in terms of nematode recorded due to the indices of similarity values that are greater or equals to

0.5. Giwa had more nematodes that were common with Unguwar Dankali and Hunkuyi while Kurmi Bomo and Basawa were two locations that have least number of nematodes in common.

For July 2013, the index of similarity between the locations (Basawa, IAR Samaru,

Giwa, Ungwa Dankali, Kurmi Bomo and Hunkuyi) and number of nematodes they have in common in root is presented in Table 12. Nematodes recorded in the root of Jatropha accessions at Basawa, IAR Samaru, Giwa, Unguwar Dankali and Kurmi-Bomo was similar. There was total dissimilarity between nematodes encounted in Hunkuyi and other locations. Fewer nematodes were recovered from Jatropha roots in Hunkuyi. IAR

Samaru and Unguwar Dankali have more nematodes in common compared with

Hunkuyi and Basawa which had the least number of nematodes in common

(dissimilarity).

For September 2013, the index of similarity between the locations (Basawa, IAR

Samaru, Giwa, Ungwar Dankali, Kurmi Bomo and Hunkuyi) and number of nematodes they have in common in soil is presented in Table 13. All locations were similar in terms of nematode recorded due to the indices of similarity values that are greater or equals to

0.5. There was a perfect similarity between Hunkuyi and IAR Samaru and Hunkuyi and

Giwa.

Table 11 Indices of similarity for plant parasitic nematode genera in soil of Jatropha stands in six location of Kaduna State, July 2013

Locations Basawa IAR Samaru Giwa Unguwar Dankali Kurmi Bomo Hunkuyi

Basawa 1.00 0.88 0.90 0.90 0.83 0.87 IAR Samaru 1.00 0.92 0.95 0.89 0.89 Giwa 1.00 0.97 0.92 0.97

Unguwar Dankali 1.00 0.95 0.94 Kurmi Bomo 1.00 0.94

Hunkuyi 1.00

IS value of 1.0 indicates total similarity between two zones, while descending values show increasing dissimilarity.

Table 12 Indices of similarity for plant-parasitic nematode genera in root of Jatropha stands in six location of Kaduna State, July 2013

Locations Basawa IAR Samaru Giwa Unguwar Dankali Kurmi Bomo Hunkuyi

Basawa 1.00 0.76 0.85 0.73 0.63 0.15 IAR Samaru 1.00 0.94 0.95 0.75 0.20 Giwa 1.00 0.89 0.80 0.22 Unguwar Dankali 1.00 0.71 0.18 Kurmi Bomo 1.00 0.25

Hunkuyi 1.00

IS value of 1.0 indicates total similarity between two zones, while descending values show increasing dissimilarity.

Table 13 Indices of similarity for plant-parasitic nematode genera in soil of Jatropha stands in six locations of Kaduna State, September 2013

Locations Basawa IAR Giwa Unguwar Dankali Kurmi Bomo Hunkuyi Samaru Basawa 1.00 0.89 0.89 0.91 0.92 0.89

IAR Samaru 1.00 1.00 0.97 0.97 1.00 Giwa 1.00 0.97 0.97 1.00 Unguwar Dankali 1.00 0.94 0.97 Kurmi Bomo 1.00 0.97

Hunkuyi 1.00 IS value of 1.0 indicates total similarity between two zones, while descending values show increasing dissimilarity.

For September 2013, the index of similarity between the locations (Basawa, IAR

Samaru, Giwa, Ungwar Dankali, Kurmi Bomo and Hunkuyi) and number of nematodes they have in common in root is presented in Table 14. The locations, IAR Samaru and

Basawa, IAR Samaru and Giwa were similar in terms of number of Nematode recorded.

There were no similarities between nematodes recorded at Kurmi Bomo and other locations. Nematodes were not encountered at Giwa and Hunkuyi and consequently the perfect dissimilarity between the two locations.

At 2 WAI, un-inoculated IARJAT2009020 gave the highest plant height (32.25) but it was not significantly higher than un-inoculated IARJAT2009041 (Table 15). Inoculated

IARJAT2009016 gave the lowest plant height (24.08) but was not significantly lower than inoculated IARJAT2009012 and IARJAT2009041. At 4 WAI, un-inoculated

IARJAT2009020 had the highest plant height (35.17) but was not significantly different from un-inoculated IARJAT2009012, IARJAT2009041 and IARJAT2009016.

Inoculated IARJAT2009041 had the lowest plant height (26.67) but was not statistically different from inoculated IARJAT2009012 and IARJAT2009016. At 6 WAI, un- inoculated IARJAT2009020 had the highest plant height (36.53) but was not statistically different from un-inoculated IARJAT2009012, IARJAT2009041 and IARJAT2009016 and inoculated IARJAT2009020. Inoculated IARJAT2009041 gave the least mean value

(29.47) and was not statistically different from inoculated IARJAT2009012and

IARJAT2009016. At 8 WAI, inoculated IARJAT2009012 had the highest mean value

(42.23) but was not significantly different from un-inoculated IARJAT2009020,

IARJAT2009012, IARJAT2009041, IARJAT2009016 and inoculated IARJAT2009020.

Inoculated IARJAT2009041 and IARJAT2009016 had the least mean value (35.62).

At 2 WAI, un-inoculated IARJAT2009020 gave the highest stem girth (0.89) but it was

Table 14 Indices of similarity for plant-parasitic nematode genera in root of Jatropha stands in six locations of Kaduna State, September 2013.

Locations Basawa IAR Samaru Giwa Unguwar Dankali Kurmi Bomo Hunkuyi Basawa 1.00 0.55 0.00 0.29 0.33 0.00 IAR Samaru 1.00 0.00 0.60 0.44 0.00 Giwa 0.00 0.00 0.00 0.00 Unguwar Dankali 1.00 0.40 0.00 Kurmi Bomo 1.00 0.00

Hunkuyi 0.00

IS value of 1.0 indicates total similarity between two zones, while descending values show increasing dissimilarity.

Table 15 Effect of Meloidogyne incognita infection on plant height of four accessions of Jatropha at Zaria, 2013

Plant Height (cm) at: Treatment 2WAI 4WAI 6WAI 8WAI

IARJAT2009020 32.25a 35.17a 36.53a 39.75ab IARJAT2009020 + Meloidogyne incognita 27.75bcd 30.33bc 33.70abc 36.95ab IARJAT2009012 28.90bc 32.47abc 34.85ab 39.87ab IARJAT2009012 + Meloidogyne incognita 25.50cde 28.83cd 32.30bcd 42.23a IARJAT2009041 30.42ab 32.67ab 34.50abc 38.75ab IARJAT2009041 + Meloidogyne incognita 24.83de 26.67d 29.47d 35.62b IARJAT2009016 28.58bc 32.35abc 33.95abc 38.75ab IARJAT2009016 + Meloidogyne incognita 24.08e 29.92cd 30.98cd 35.62b

SE± 0.966 1.007 1.038 1.642

Mean in a column followed by different letters are significantly different using DMRT at 5% level of significance.

WAI = Weeks After Inoculation

not significantly higher than un-inoculated IARJAT2009012, IARJAT2009041 and

IARJAT2009016 (Table 16). Inoculated IARJAT2009016 gave the lowest stem girth

(0.70) but was not significantly different from un-inoculated IARJAT2009012, inoculated IARJAT2009020, IARJAT2009012 and IARJAT2009041. At 4 WAI, un- inoculated IARJAT2009020 had the highest stem girth (1.15) but was not significantly different from un-inoculated IARJAT2009041 and IARJAT2009016. Inoculated

IARJAT2009041 had the lowest stem girth (0.94) but was not significantly different from un-inoculated IARJAT2009012, IARJAT2009041, inoculated IARJAT2009020,

IARJAT2009012 and IARJAT2009016. At 6 WAI, un-inoculated IARJAT2009020 had the highest stem girth (1.39) but was not statistically different from un-inoculated

IARJAT2009012, IARJAT2009041 and IARJAT2009016. Inoculated IARJAT2009041 gave the least mean value (1.10) and was not significantly different from un-inoculated

IARJAT2009012, IARJAT2009041, inoculated IARJAT2009020, IARJAT2009012 and

IARJAT2009016. At 8 WAI, inoculated IARJAT2009020 had the highest mean value

(1.82) but was not significantly different from un-inoculated IARJAT2009012,

IARJAT2009041, IARJAT2009016 and inoculated IARJAT2009012. Inoculated

IARJAT2009020and IARJAT2009041 had the least stem girth (1.30) but was not significantly different from un-inoculated IARJAT2009012, IARJAT2009041,

IARJAT2009016, IARJAT2009012 and IARJAT2009016

At 2 WAI, un-inoculated IARJAT2009020and IARJAT2009012gave the highest numbers of leaf (6.67) but it was not significantly higher than un-inoculated

IARJAT2009041,inoculated IARJAT2009020 and IARJAT2009012 (Table 17). Un- inoculated IARJAT2009016 gave the lowest number of leaf (5.50) but was not significantly different from un-inoculated IARJAT2009041, inoculated

IARJAT2009020, IARJAT2009012, IARJAT2009041 and IARJAT2009016. At 4 WAI,

Table 16 Effect of Meloidogyne incognita infection on stem girth of four accessions of Jatropha

Stem Girth (cm) at: Treatment 2WAI 4WAI 6WAI 8WAI IARJAT2009020 0.89a 1.15a 1.39a 1.82a

IARJAT2009020 + Meloidogyne incognita 0.75bcd 0.96b 1.13c 1.30b IARJAT2009012 0.78abdc 1.00b 1.25abc 1.58ab IARJAT2009012 + Meloidogyne incognita 0.71dc 0.97b 1.18bc 1.57ab IARJAT2009041 0.83abc 1.05ab 1.24abc 1.56ab IARJAT2009041 + Meloidogyne incognita 0.72dc 0.94b 1.10c 1.30b IARJAT2009016 0.85ab 1.14a 1.32ab 1.58ab

IARJAT2009016 + Meloidogyne incognita 0.70d 0.95b 1.18bc 1.37b SE± 0.036 0.037 0.049 0.079

Mean in a column followed by different letters are significantly different using DMRT at 5% level of significance.

WAI = Weeks After Inoculation

Table 17 Effect of Meloidogyne incognita infection on numbers of leaf counted on Jatropha plants

Leaf Number at: Treatment 2WAI 4WAI 6WAI 8WAI IARJAT2009020 6.67a 8.33a 11.33a 14.17ab

IARJAT2009020 + Meloidogyne incognita 6.17ab 7.50b 9.17ab 10.17b IARJAT2009012 6.67a 8.50ab 11.50a 13.50ab IARJAT2009012 + Meloidogyne incognita 6.00ab 7.33b 11.17a 14.50a IARJAT2009041 6.17ab 8.00ab 9.67ab 12.33ab IARJAT2009041 + Meloidogyne incognita 5.67b 5.83c 8.33b 10.83ab IARJAT2009016 5.50b 7.67ab 11.00a 14.00ab

IARJAT2009016 + Meloidogyne incognita 5.67b 8.17ab 11.00a 13.33ab

SE± 0.227 0.354 0.696 1.076

Mean in a column followed by different letters are significantly different using DMRT at 5% level of significance.

WAI = Weeks After Inoculation

un-inoculated IARJAT2009020 had the highest number of leaf (8.33) but was not significantly different from un-inoculated IARJAT2009012, IARJAT2009041 and

IARJAT2009016. Inoculated IARJAT2009041 had the lowest number of leaf (5.83) but was significantly different from other treatments. At 6 WAI, un-inoculated

IARJAT2009020 had the highest number of leaf (11.33) and was statistically different from inoculated IARJAT2009041 that had the least (8.33) numbers of leaf. At 8 WAI, inoculated IARJAT2009012 had the highest mean value (14.50) and was significantly different from inoculated IARJAT2009020 that had the least number of leaves (10.17).

Table 18 shows the gall index and the nematode population on Jatropha curcas accessions and tomato plants inoculated with Meloidogyne incognita. The population of

M. incognita differed between the tomato variety (Roma VF) which was used as standard check and the Jatropha curcas accessions but there was no significant difference between the Jatropha curcas accessions. The greatest reduction in number of galls (3.17) was observed in IARJAT2009012 accession of Jatropha followed by

IARJAT2009020 (3.50) and IARJAT2009041 (6.67). The reduction in number of eggs and final nematode population obtained shows that IARJAT2009041 has the highest reduction, followed by IARJAT2009020 and IARJAT2009016. There were significant difference between the controls and standard check on the number of M. incognita eggs and nematode population extracted after 8 weeks of inoculation from the root of inoculated plants and the soil. There was no significant different in the mean obtained for all the Jatropha accessions. Roma VF used as standard check has the highest number of nematode population (954.53).

Table 18 Number of galls, population of Meloidogyne incognita and reproduction factor of inoculated Jatropha and tomato

Number of Galls Nematode Population Reproduction Factor Treatments

IARJAT2009020 + Meloidogyne incognita 3.50b 18.77b 0.01

IARJAT2009012 + Meloidogyne incognita 3.17b 37.27b 0.02

IARJAT2009041 + Meloidogyne incognita 6.67b 13.50b 0.01

IARJAT2009016 + Meloidogyne incognita 10.00b 28.71b 0.01

Roma VF + Meloidogyne incognita 387.17a 954.53a 0.48 LSD 26.269 28.357 0.014

Means in a column followed by different letters are significantly different using LSD at 5% level of significance

WAI = Weeks After Inoculation

CHAPTER FIVE

5.0 DISCUSSION

The conducted survey indicated that twenty-four (24) genera of plant parasitic nematodes were associated with Jatropha plants in four Local Government Areas

(Sabon Gari, Giwa, Kudan and Zaria) of Kaduna State. These are Scutellonema,

Hoplolaimus, Pratylenchus, Aphelenchus, Meloidogyne, Tylenchoryhnchus,

Rotylenchus, Longidorus, Helicotylenchus, Paratylenchus, Heterodera , Xiphinema,

Tylenchus, Criconemoides, Hemicycliophora, Aphelenchoides, Tetylenchus,

Trichodorus, Dorylaimus, Tylenchulus, Telotylenchus, Pratylenchoides,

Telotylenchoides and Rotylenchoides. All the twenty-four genera of plant parasitic nematodes were recorded in soil of Jatropha in July 2013; however, only eleven genera of the nematodes were recorded from the root of Jatropha in the same survey. These are

Scutellonema, Hoplolaimus, Pratylenchus, Meloidogyne, Rotylenchus, Longidorus,

Helicotylenchus, Paratylenchus Heterodera, Xiphinema and Aphelenchoides Also, twenty (20) genera of plant parasitic nematodes were recorded in the soil and six from the root of Jatropha during September 2013 survey. Thus, those not found associated with Jatropha in September are Telotylenchus, Pratylenchoides, Telotylenchoides and

Rotylenchoides. Emeh, (2012), recorded seventeen (17) genera of plant parasitic nematodes to be associated with Jatropha curcas accessions in Zaria. These comprise all the genera reported in this study except Aphelenchus, Paratylenchus, Heterodera,

Aphelenchoides, Tetylenchus, Dorylaimus, Tylenchulus, Telotylenchus, Pratylenchoides,

Telotylenchoides and Rotylenchoides. The increase in nematode genera due to these findings may be increase in number of sampling locations, change in time and Jatropha accessions.

Scutellonema was the most abundant plant parasitic nematode found on Jatropha curcas

in all the fields. It was recovered from the soil as well as from the roots. This finding corroborates the findings of Emeh, 2012, that Scutellonema spp. had the highest population

The frequency of occurrence and prominence value of Meloidogyne in this study corroborate Begum (1996) report that Meloidogyne spp. may be important on Jatropha.

Xiphinema, Longidorus and Trichodorus were recorded in this study. Mekete (2010) reported the association of these nematode species among others with biofuel crops. The importance of these nematodes as vectors of plant viral diseases is well documented

(Brown et. al., 1993). Kashina et al. (2013) reported the occurrence of begomovirus in

Jatropha curcas, the frequenct association of these nematodes in this study asserts that they may be vectoring this disease.

Generally, more plant parasitic nematodes were recorded in the soil and root of Jatropha in July, 2013 than in of September, 2013. The values reported for the indices of similarity in the soil and roots indicate that there are more similarities in number of nematodes recorded in July, 2013 than September, 2013. This might be attributed to the fact that plant parasitic nematode population will be greater with the abundance of soil moisture (Agrios, 2005). Therefore decline in nematode population in the September compared to July may be due to difference in the amount of soil moisture between July and September, 2013. In orchards, Jatropha fields were constantly clear of unwanted plants, fertilizers were applied and watering of plants was done. The plants were well taken care of whereas no such treatments were given to Jatropha plants on the farmer‟s field. The higher number of nematodes found in the Orchards compared to the farmer‟s field might be due to these management practices that may tend to favour population build up in orchards than other farms.

Based on the index of similarity values across all locations, nematode genera encountered are similar. Differences observed in plant parasitic nematode communities where it exists can be attributed primarily to the different management practices in each of the location which in turn disturb soil nematode community structure and function

(Neher, 1995; Neher and Cambell, 1994).

The pathogenicity test shows that Meloidogyne incognita did not significantly reduce the growth of all the Jatropha accessions used as there were no significant differences in the growth parameters of both inoculated and un-inoculated accessions. Meloidogyne incognita reproduced maximally on tomato which was used as standard check but maximum reproduction was hindered in the roots of Jatropha accessions. Inability of

Meloidogyne incognita to reproduce well on the Jatropha may be due to pest and pathogens repellant toxins described by Heller (1996). The Jatropha accessions tested appears to be tolerant to Meloidogyne incognita infections. Begum (1996) reported that

Jatropha podagrica is a parasite of Meloidogyne incognita. The apparent discrepancy between the findings in this study and that of Begum (1996) may be due to differences in varieties and the Meloidogyne species. It is possible that the accessions used in this study are fairly resistant to Meloidogyne incognita. There is therefore the need to screen more cultivars against the nematode, Meloidogyne incognita.

CHAPTER SIX

SUMMARY, CONCLUSION AND RECOMMENDATIONS

6.1 Summary

A study was conducted to determine the genera, frequency and prominence value of plant parasitic nematodes associated with Jatropha curcas in Sabon Gari, Kudan, Giwa and Zaria Local Government areas of Kaduna State, Nigeria. Twenty-four genera of plant parasitic nematodes were recorded in all the locations. Plant parasitic nematodes recovered include Scutellonema, Hoplolaimus, Pratylenchus, Aphelenchus,

Meloidogyne, Tylenchoryhnchus, Rotylenchus, Longidorus, Helicotylenchus,

Paratylenchus Heterodera Xiphinema, Tylenchus, Criconemoides, Hemicycliophora,

Aphelenchoides, Tetylenchus, Trichodorus, Dorylaimus, Tylenchulus, Telotylenchus,

Pratylenchoides, Telotylenchoides, and Rotylenchoides. The most prominent nematode from the soil were Scutellonema with prominence value of 81.09, followed by

Meloidogyne and Rotylenchus with prominence values of 46.50 and 39.60, respectively and those from the roots were Scutellonema, Meloidogyne and Rotylenchus with prominence values of 77.15, 50.93 and 26.94, respectively

Screen house experiment was conducted to determine the pathogenicity of Meloidogyne incognita, on four Jatropha curcas accessions. Data on plant height, stem girth and number of leaves together with gall index and nematode reproduction were collected.

Evaluation of four accessions of Jatropha curcas (IARJAT2009020, IARJAT2009011,

IARJAT2009041 and IARJAT2009016) shows that these Jatropha accessions are resistant to Meloidogyne incognita.

6.2 Conclusion

The study has demonstrated that twenty-four genera of plant parasitic nematodes are

associated with Jatropha curcas accessions in Sabon Gari, Kudan, Giwa and Zaria

Local Government areas of Kaduna State, Nigeria. IARJAT2009020, IARJAT2009011,

IARJAT2009041 and IARJAT2009016 prove to be resistance to Meloidogyne incognita.

It can therefore be concluded that although plant parasitic nematodes were recorded in the soil and root of Jatropha in sampled areas, Meloidogyne incognita is not pathogenic on Jatropha curcas accessions used. IARJAT2009020, IARJAT2009011,

IARJAT2009041 and IARJAT2009016 can therefore be used in mixed crop combinations to manage Meloidogyne incognita infected soils.

6.3 Recommendations

Based on the findings of this study,

1. Further pathogenicity test of other important nematodes to the harvest of the

crop need to be determined in order to ascertain the damage caused by nematode

that will translate to yield loss of Jatropha.

2. Screening of more accessions of Jatropha against Meloidogyne incognita and

other prominent nematodes should be carried out.

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Appendix ANOVA Tables

ANOVA Effect of Meloidogyne incognita infection on plant height of four accessions of Jatropha

2 Weeks After Inoculation

Source DF Sum of Squares Mean Square F Value Pr > F

Model (Accessions * Treatments) 7 338.2764583 48.3252083 6.48 <.0001

Error 40 298.3083333 7.4577083

Total 47 636.5847917

4 Weeks After Inoculation

Source DF Sum of Squares Mean Square F Value Pr > F

Model (Accessions * Treatments) 7 313.9466667 44.8495238 5.53 0.0002

Error 40 324.2833333 8.1070833

Total 47 638.23

6 Weeks After Inoculation

Source DF Sum of Squares Mean Square F Value Pr > F

Model (Accessions * Treatments) 7 215.6347917 30.8049702 3.57 0.0045

Error 40 344.885 8.622125

Total 47 560.5197917

8 Weeks After Inoculation

Source DF Sum of Squares Mean Square FValue Pr > F

Model (Accessions * Treatments) 7 214.558392 30.651199 1.42 0.2237

Error 40 862.300333 21.557508

Total 47 1076.858725

ANOVA 14 Effect of Meloidogyne incognita infection on stem girth of four accessions of Jatropha

2 Weeks After Inoculation

Source DF Sum of Squares Mean Square F Value Pr > F

Model (Accessions * Treatments) 7 0.22 0.03142857 3.07 0.011

Error 40 0.40916667 0.01022917

Total 47 0.62916667

4 Weeks After Inoculation

Source DF Sum of Squares Mean Square F Value Pr > F

Model (Accessions * Treatments) 7 0.30411458 0.04344494 3.87 0.0027

Error 40 0.44958333 0.01123958

Total 47 0.75369792

6 Weeks After Inoculation

Source DF Sum of Squares Mean Square F Value Pr > F

Model (Accessions * Treatments) 7 0.45286458 0.06469494 3.39 0.0062

Error 40 0.76291667 0.01907292

Total 47 1.21578125

8 Weeks After Inoculation

Source DF Sum of Squares Mean Square F Value Pr > F

Model (Accessions * Treatments) 7 1.31453125 0.18779018 3.71 0.0035

Error 40 2.02375 0.05059375

Total 47 3.33828125

ANOVA Effect of Meloidogyne incognita infection on numbers of leaf counted on four accessions of Jatropha

2 Weeks After Inoculation

Source DF Sum of Squares Mean Square F Value Pr > F

Model (Accessions * Treatments) 7 8.3125 1.1875 2.88 0.0156

Error 40 16.5 0.4125

Total 47 24.8125

4 Weeks After Inoculation

Source DF Sum of Squares Mean Square F Value Pr > F

Model (Accessions * Treatments) 7 35.3125 5.04464286 5.02 0.0004

Error 40 40.16666667 1.00416667

Total 47 75.47916667

6 Weeks After Inoculation

Source DF Sum of Squares Mean Square F Value Pr > F

Model (Accessions * Treatments) 7 58.3125 8.3303571 2.15 0.0603

Error 40 155.1666667 3.8791667

Total 47 213.4791667

8 Weeks After Inoculation

Source DF Sum of Squares Mean Square F Value Pr > F

Model (Accessions * Treatments) 7 107.8125 15.4017857 1.66 0.1458

Error 40 370.1666667 9.2541667

Total 47 477.9791667

ANOVA Table for Gall number of inoculated Meloidogyne incognita Jatropha accessions and Tomato (Roma VF).

Number of Galls

Source DF Sum of Squares Mean Square F Value Pr > F

Model (Accessions * Treatments) 4 698176.2000 174544.0500 42.16 0.0001

Error 25 103510.5000 4140.4200

Total 29 801686.7000

ANOVA Table for Nematode Population of inoculated Meloidogyne incognita Jatropha accessions and Tomato (Roma VF).

Nematodes Population

Source DF Sum of Squares Mean Square F Value Pr > F

Model (Accessions * Treatments) 4 4153278.902 1038319726 215.22 0.0001

Error 25 120613.841 4824.554

Total 29 4273892